Comparison of different electroporation parameters on transfection efciency of sheep testicular cells

Objective: Electroporation can be a highly efficient method for introducing the foreign genetic materials into the targeted cells for transient and/or permanent genetic modification. Considering the application of this technique as a very efficient method for drug, oligonucleotide, antibody and plasmid delivery for clinical applications and production of transgenic animals, the present study aimed to optimize the transfection efficiency of sheep testicular cells including spermatogonial stem cells (SSCs) via electroporation. Materials and Methods: This study is an experimental research conducted in Biotechnology Research Center (Avicenna Research Institute, Tehran, Iran) from September 2013 to March 2014. Following isolation and propagation of one-month lamb testicular cells (SSCs and somatic testicular cells including; Sertoli, Leydig, and myoid cells), the effect of different electroporation parameters including total voltages (280, 320, and 350 V), burst durations (10, 8, and 5 milliseconds), burst modes (single or double) and addition of dimethyl sulfoxide (DMSO) were evaluated on transfection efficiency, viability rate and mean fluorescent intensity (MFI) of sheep testicular cells. Results: The most transfection effciency was obtained in 320 V/8 milliseconds/single burst group in transduction medium with and without DMSO. There was a signifcantly inverse correlation between transfection effciency with application of both following parameters: addition of DMSO and double burst. After transfection, the highest and lowest viability rates of testicular cells were demonstrated in 320 V/8 milliseconds with transduction medium without DMSO and 350 V/5 milliseconds in medium containing DMSO. Addition of DMSO to transduction medium in all groups signifcantly decreased the viability rate. The comparison of gene expression indicated that Sertoli and SSCs had the most?uorescence intensity in 320 V/double burst/DMSO positive. However, myoid and Leydig cells showed the maximum expression in 320 V/single burst and/or 350 V/double burst/DMSO positive. Conclusion: We optimized the electroporation method for transfection of sheep testicular cells and recommended the application of 320 V/8 milliseconds/single pulse/DMSO negative for transduction of plasmid vector into these cells. Among testicular cells, the most external gene expression was demonstrated in SSC population

Anatomy, histology and histochemistry of accessory sex glands in male Persian squirrel (Sciurus anomalus)

Persian squirrel (Sciurus anomalus) is a species of squirrels in the Middle East. There is little information about the anatomy and histology of various organs of this species. Moreover, there are no practical data about the accessory male sex glands of this species of squirrel. In this study the anatomical, histological and histochemical properties of the male reproductive system accessory glands of Persian Squirrel was evaluated. Eight adult male squirrels were anesthetized and euthanized. The pelvic area was dissected and the male reproductive system was separated. The accessory sex glands were investigated for gross anatomical aspect. Samples were fixed in formaldehyde for histological and histochemical studies. The coagulating glands, prostate and bulbourethral glands were observed at gross anatomical level. A single heart-shape and compact prostate gland was situated on the dorsal side of pelvic urethra. Two small coagulating glands were observed on the cranio-dorsal side of prostate. Two large spiral shape bulbourethral glands were situated out of the pelvic cavity near the root of penis on both sides of anal area. Histologic studies revealed that all accessory sex glands were alveolar glands with cuboidal to columnar epithelium. The most positive reaction for PAS stain was observed in the trabeculae of glands. There was no positive reaction for lipids in glands stained with Oil red O and Sudan Black. Some anatomical differences were observed in the accessory sex glands of Persian Squirrel in comparison to other rodents, however the histology of glands was almost similar to other rodents

Effect of monosodium glutamate on testicular tissue of paclitaxel-treated mice: an experimental study

Background: Paclitaxel (PTX), a chemotherapeutic agent, and monosodium glutamate (MSG) have oxidative effects on testicular tissue. Objective: In this study, the effects of MSG administration on the exacerbation of testicular tissue alterations related to PTX treatment were evaluated. Materials and Methods: MSG (30 & 60 mg/kg i.p.) was administrated to six groups (n = 8/each) of adult mice before or after PTX treatment: control, PTX-treated, MSG30 + PTX, MSG60 + PTX, PTX + MSG30, and PTX + MSG60. Following the euthanizing, the body weight measurement, pituitary–testicular axis hormonal analysis and serum lipid peroxidation index assessment was prepared, testicular histomorphometry (tubular diameter and germinal epithelium height), immunohistochemistry of p53 was completed. Microscopic indices of spermatogenesis (tubular differentiation, spermiogenesis and repopulation indices) were studied. Results: Body weight was not changed significantly. The levels of testosterone (p = 0.0001), follicle stimulating hormone (p = 0.019), and luteinizing hormone (p = 0.08) were decreased while the level of lipid peroxidation index was increased (p = 0.208) in the treated groups. The histomorphometry indices (p < 0.0001 and p = 0.001, respectively), germ cells population (p < 0.05) and microscopic indices of spermatogenesis (p = 0.001, p = 0.005, p < 0.0001, respectively) were significantly reduced in all treated groups. The administration of MSG before PTX treatment induces more changes. The most positive reaction to p53 was observed in MSG30 or 60 + PTX groups compared to other groups. Conclusion: The administration of MSG could intensify testicular tissue alterations related to PTX chemotherapy. Key words: Mice, Monosodium glutamate, Morphometry, Paclitaxel, Testicular tissue

Evaluation of Possible Effects of Hyoscine in Xylazine-Induced Fetal Death in Pregnant Rats

Although xylazine is widely used in domestic animals as a sedative, analgesic, and muscle relaxant, its side effects on the uterus prevent its utilization in pregnant animals or in embryo transfer. Although the effects of xylazine on increasing uterine contractions have been confirmed, no reliable report of fetal death due to xylazine administration has been published. Hyoscine is an anticholinergic medication that has antimuscarinic and antispasmodic effects in the uterine tissue of pregnant cattle during in vitro studies, therefore, we investigated if administration of xylazine in the last third of pregnancy could increase fetal death and if hyoscine could prevent its adverse effects. Twenty adult female rats, after mating with four adult male rats and confirming pregnancy, were randomly divided into two equal control and treatment groups. On the 18th day of pregnancy, the number of fetuses per rat was determined using ultrasonography. Rats in the treatment group received hyoscine (1 mg/kg, intraperitoneally) for 3 days. Subsequently, all rats were administered xylazine (10 mg/kg, intraperitoneally) for 3 days. On the 21st day of pregnancy, the number of living and dead fetuses was counted after laparotomy. Also, the weight and dimensions of the fetuses were measured. The results showed that although more fetuses lost their lives in the treatment group compared to the control group, the statistical difference in the percentage of fetal mortality in the two groups was not significant (p 0.05). In addition, the comparison of the mean weight, body length, and body width of living and dead fetuses in both groups showed that there was no statistically significant difference between these groups (p 0.05). It could be concluded that maternal xylazine intake in rats could cause about 18-25% of fetal mortality. However, the use of hyoscine to prevent fetal death induced by xylazine is not recommended

The association between fat mass and obesity‐associated ( FTO ) genotype and serum vitamin D level in breast cancer patients

Abstract: The preventive effect of vitamin D against breast cancer can be influenced by gene polymorphisms. This study aimed to investigate the association between serum level of 25(OH) vitamin D and FTO genotype in breast cancer patients. A cross‐sectional study was carried out on 180 newly diagnosed patients with breast cancer in Tehran, Iran. The blood samples were collected from the participants in order to assess the FTO gene rs9939609 polymorphism by the tetra‐primer amplification refractory mutation system (Tetra‐ARMS) PCR method. The serum level of 25(OH) vitamin D was measured using the direct competitive enzyme‐linked immunosorbent assay (ELISA) method. The association between vitamin D and the FTO genotype in patients with breast cancer was assessed after adjustment for cofounders. The frequency of TT, AT and AA genotypes in the breast cancer patients were 43% (n = 77), 49% (n = 89) and 8% (n = 14), respectively. All patients with higher than 40 ng/dl of serum 25(OH) vitamin D had one or two copies of FTO rs9939609 risk allele (p = 0.019). No linear association was found between the number of FTO risk allele and the level of serum vitamin D. All patients with high serum level of 25(OH) vitamin D had one or two copies of FTO rs9939609 risk allele. FTO gene polymorphisms may counteract the beneficial effects of vitamin D in breast cancer prevention. Further studies can help to better understand the genetic factors predisposing to breast cancer and their effect on the association between vitamin D and breast cancer

The Association of Fat-Mass-and Obesity-Associated Gene Polymorphism (rs9939609) With Colorectal Cancer: A Case-Control Study

Background and Aim: The association between the rs9939609 polymorphism of fat mass and obesity-associated gene (FTO) and risk of colorectal cancer is controversial. This study aims to evaluate the relationship between FTO rs9939609 polymorphism and colorectal cancer (CRC) in Iranian people. Methods: A case-control study was conducted on 125 patients with CRC and 250 healthy subjects in Tehran, Iran. Demographic data and blood samples were collected from all participants. Genotyping of rs9939609 polymorphism was performed by the tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) method. Results: The occurrence of AA genotype of FTO rs9939609 polymorphism in the colorectal cancer patients was significantly higher compared to that of healthy subjects (16.4 vs. 2.9%, respectively, P=0.02). The association between the frequency of risk allele of the FTO polymorphism and CRC (B=1.67, P=0.042) remained significant after adjustment for age. Further adjustment for gender (model 2) and marital status (model 3) did not change this result (B=1.67, P= 0.042 and B=1.67, P=0.043, respectively). The results remained significant after additional adjustment for ethnicity (B=1.57, P= 0.047). Conclusion: We found a positive association between the A allele of the rs9939609 polymorphism and CRC. Future studies are required to identify the underlying mechanisms

ILSF, A THIRD GENERATION LIGHT SOURCE LABORATORY IN IRAN

Abstract The Iranian Light Source Facility (ILSF) project is a first large scale accelerator facility which is currently under planning in Iran. On the basis of the present design, circumference of the 3 GeV storage ring is 297.6 m. Beam current and natural beam emittance are 400 mA and 3.278 nm.rad respectively. The facility will be built on a land of 50 hectares area in the city of Qazvin, located 150 km West of Tehran. The city is surrounded by many universities, research centers and industrial companies. The design and construction of prototype items such as radio frequency solid state amplifier, dipole magnets, highly stable magnet power supplies and girders have already begun. Site selection studies, including geotechnical and seismological measurements are being performed. Conceptual Design Report, CDR, as the first milestone of the project was published in October 2012